Moskaitis J E, Sargent T D, Smith L H, Pastori R L, Schoenberg D R
Department of Pharmacology, Uniformed Services University of the Health Sciences, Bethesda, Maryland 20814-4799.
Mol Endocrinol. 1989 Mar;3(3):464-73. doi: 10.1210/mend-3-3-464.
In adult Xenopus serum, albumin gene expression is regulated by estrogen through the selective destabilization of its mRNA during the vitellogenic response. The present study reports the cDNA sequence of both the 68K and 74K Xenopus albumin mRNAs, their derived amino acid sequence, and the regulation of albumin gene expression during embryogenesis. Albumin mRNA has a 39 nucleotide 5' untranslated region terminating in a consensus translation initiation site. The derived amino acid sequence yields a 24-amino acid hydrophobic leader sequence (terminating in Lys-Arg) that shares significant homology with the leader peptide of rat albumin. Overall there is 37% sequence identity between rat and frog albumin, with exact conservation of all but one Cys residue and the Pro residues responsible for the three domain structure of the mature protein. The 74K albumin (unlike the 68K albumin) is glycosylated; a point mutation converting Lys256 to Asn introduces an N-linked glycosylation site that is similar to one found in the sequence of mammalian alpha-fetoproteins. A larval albumin-like protein was not detectable by silver staining in serum of tadpoles before the beginning of metamorphosis at stage 48. Albumin mRNA is absent from early tadpoles (stages 22-47); however, it is rapidly induced at stage 48 as one of the earliest manifestations of metamorphosis. Exposure of embryos to 10(-8) M T3, which regulates amphibian metamorphosis, resulted in the premature induction of albumin mRNA, such that it is evident by stage 43.
在成年非洲爪蟾血清中,卵黄生成反应期间,雌激素通过选择性地使白蛋白mRNA不稳定来调节白蛋白基因的表达。本研究报告了68K和74K非洲爪蟾白蛋白mRNA的cDNA序列、其推导的氨基酸序列,以及胚胎发育过程中白蛋白基因表达的调控情况。白蛋白mRNA具有一个39个核苷酸的5'非翻译区,终止于一个共有翻译起始位点。推导的氨基酸序列产生一个24个氨基酸的疏水前导序列(以Lys-Arg结尾),该序列与大鼠白蛋白的前导肽具有显著同源性。总体而言,大鼠和青蛙白蛋白之间有37%的序列同一性,除了一个半胱氨酸残基和负责成熟蛋白三结构域结构的脯氨酸残基外,所有其他残基都完全保守。74K白蛋白(与68K白蛋白不同)是糖基化的;将Lys256突变为Asn的点突变引入了一个N-连接糖基化位点,该位点类似于在哺乳动物甲胎蛋白序列中发现的一个位点。在变态开始前的第48阶段,通过银染在蝌蚪血清中未检测到幼虫白蛋白样蛋白。早期蝌蚪(第22-47阶段)中不存在白蛋白mRNA;然而,它在第48阶段作为变态的最早表现之一迅速被诱导。将胚胎暴露于调节两栖动物变态的10(-8) M T3中,导致白蛋白mRNA的过早诱导,以至于在第43阶段就很明显。